1. Targeting the Myofibroblastic Cancer-Associated Fibroblast Phenotype Through Inhibition of NOX4.
- Author
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Hanley CJ, Mellone M, Ford K, Thirdborough SM, Mellows T, Frampton SJ, Smith DM, Harden E, Szyndralewiez C, Bullock M, Noble F, Moutasim KA, King EV, Vijayanand P, Mirnezami AH, Underwood TJ, Ottensmeier CH, and Thomas GJ
- Subjects
- Actins analysis, Adenocarcinoma chemistry, Adenocarcinoma genetics, Adult, Aged, Aged, 80 and over, Animals, Cancer-Associated Fibroblasts chemistry, Cancer-Associated Fibroblasts physiology, Carcinoma, Non-Small-Cell Lung chemistry, Carcinoma, Non-Small-Cell Lung genetics, Carcinoma, Squamous Cell chemistry, Carcinoma, Squamous Cell genetics, Cell Count, Cell Transdifferentiation drug effects, Cell Transdifferentiation genetics, Colorectal Neoplasms pathology, Disease Progression, Esophageal Neoplasms chemistry, Esophageal Neoplasms genetics, Female, Head and Neck Neoplasms chemistry, Head and Neck Neoplasms drug therapy, Head and Neck Neoplasms genetics, Humans, Lung Neoplasms chemistry, Lung Neoplasms genetics, Male, Mice, Middle Aged, Mouth Neoplasms pathology, Myofibroblasts chemistry, NADPH Oxidase 4, NADPH Oxidases analysis, NADPH Oxidases genetics, Neoplasm Transplantation, Oropharyngeal Neoplasms pathology, Phenotype, Pyrazoles therapeutic use, Pyrazolones, Pyridines therapeutic use, Pyridones, RNA Interference, Reactive Oxygen Species metabolism, Survival Rate, Up-Regulation, Adenocarcinoma drug therapy, Cancer-Associated Fibroblasts pathology, Carcinoma, Non-Small-Cell Lung drug therapy, Carcinoma, Squamous Cell drug therapy, Colorectal Neoplasms chemistry, Esophageal Neoplasms drug therapy, Lung Neoplasms drug therapy, Mouth Neoplasms chemistry, Myofibroblasts pathology, NADPH Oxidases antagonists & inhibitors, Oropharyngeal Neoplasms chemistry
- Abstract
Background: Cancer-associated fibroblasts (CAFs) are tumor-promoting and correlate with poor survival in many cancers, which has led to their emergence as potential therapeutic targets. However, effective methods to manipulate these cells clinically have yet to be developed., Methods: CAF accumulation and prognostic significance in head and neck cancer (oral, n = 260; oropharyngeal, n = 271), and colorectal cancer (n = 56) was analyzed using immunohistochemistry. Mechanisms regulating fibroblast-to-myofibroblast transdifferentiation were investigated in vitro using RNA interference/pharmacological inhibitors followed by polymerase chain reaction (PCR), immunoblotting, immunofluorescence, and functional assays. RNA sequencing/bioinformatics and immunohistochemistry were used to analyze NAD(P)H Oxidase-4 (NOX4) expression in different human tumors. NOX4's role in CAF-mediated tumor progression was assessed in vitro, using CAFs from multiple tissues in Transwell and organotypic culture assays, and in vivo, using xenograft (n = 9-15 per group) and isograft (n = 6 per group) tumor models. All statistical tests were two-sided., Results: Patients with moderate/high levels of myofibroblastic-CAF had a statistically significant decrease in cancer-specific survival rates in each cancer type analyzed (hazard ratios [HRs] = 1.69-7.25, 95% confidence intervals [CIs] = 1.11 to 31.30, log-rank P ≤ .01). Fibroblast-to-myofibroblast transdifferentiation was dependent on a delayed phase of intracellular reactive oxygen species, generated by NOX4, across different anatomical sites and differentiation stimuli. A statistically significant upregulation of NOX4 expression was found in multiple human cancers (P < .001), strongly correlating with myofibroblastic-CAFs (r = 0.65-0.91, adjusted P < .001). Genetic/pharmacological inhibition of NOX4 was found to revert the myofibroblastic-CAF phenotype ex vivo (54.3% decrease in α-smooth muscle actin [α-SMA], 95% CI = 10.6% to 80.9%, P = .009), prevent myofibroblastic-CAF accumulation in vivo (53.2%-79.0% decrease in α-SMA across different models, P ≤ .02) and slow tumor growth (30.6%-64.0% decrease across different models, P ≤ .04)., Conclusions: These data suggest that pharmacological inhibition of NOX4 may have broad applicability for stromal targeting across cancer types., (© The Author 2017. Published by Oxford University Press.)
- Published
- 2018
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